Papilloma viruses, products for the detection thereof as well as for treating diseases caused by them

ABSTRACT

This invention relates to a DNA coding for a peptide of a papilloma virus major capsid protein and a papilloma virus genome, respectively. Furthermore, this invention concerns proteins coded by the papilloma virus genome and antibodies directed thereagainst as well as the use thereof for diagnosis, treatment and vaccination.

This is a national phase filing of the Application No. PCT/DE97/02659,which was filed with the Patent Corporation Treaty on Nov. 12, 1997, andis entitled to priority of the German Patent Application DE 196 48962.8, filed Nov. 26, 1996.

I. FIELD OF THE INVENTION

This invention relates to a DNA coding for a peptide of a papillomavirus major capsid protein and a papilloma virus genome, respectively.In addition, this invention concerns proteins coded by the papillomavirus genome and antibodies directed against them as well as their usefor diagnosis, treatment and vaccination.

II. BACKGROUND OF THE INVENTION

It is known that papilloma viruses infect the epithelium of human beingsand animals. Human papilloma viruses (hereinafter referred to as HPviruses) are found in benign epithelial neoplasms, e.g. warts,condylomas in the genital zone, and malignant epithelial neoplasms, e.g.carcinomas of the skin and the uterus (zur Hausen, 1996, Biochimica etBiophysica Acta (BBA) 1288:55-78). HP viruses are also considered forthe growth of malignant tumors in the oropharyngeal zone (zur Hausen,1977, Curr. Top. Microbiol. Immunol. 78:1-30).

Papilloma viruses have an icosahedral capsid without envelope in which acircular, double-stranded DNA molecule of about 7900 bp is present. Thecapsid comprises a major capsid protein (L1) and a minor capsid protein(L2). Both proteins, coexpressed or L1 expressed alone, result in vitroin the formation of virus-like particles (Kirnbauer et al., 1993,Journal of Virology 67:6929-6936).

Papilloma viruses cannot be proliferated in monolayer cell culture.Therefore, their characterization is extremely difficult, the detectionof papilloma viruses already creating considerable problems. Thisapplies especially to papilloma viruses in carcinomas of the skin.

Thus, it is the object of the present invention to provide a product bywhich papilloma viruses can be detected, particularly in carcinomas ofthe skin. Furthermore, a product should be provided to be able to taketherapeutic steps against these papilloma viruses.

According to the invention, this is achieved by providing the subjectmatters in the claims.

III. SUMMARY OF THE INVENTION

This invention relates to a DNA coding for a peptide of a papillomavirus major capsid protein and a papilloma virus genome, respectively.Furthermore, this invention concerns proteins coded by the papillomavirus genome and antibodies directed thereagainst as well as the usethereof for diagnosis, treatment and vaccination.

IV. BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the base sequence (SEQ ID NOS:1 and 3) and the amino acidsequence (SEQ ID NO:2), derived therefrom, of a DNA coding for a peptideof L1 of a papilloma virus. This DNA was deposited as plasmid DL17 withDSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen (German-typecollection of micro-organisms and cell cultures)) under DSM 11180 onSep. 17, 1996.

FIG. 2 shows the base sequence (SEQ ID NOS:4 and 6) and the amino acidsequence (SEQ ID NO:5), derived therefrom, of a DNA coding for a peptideof L1 of a papilloma virus. This DNA was deposited as plasmid DL20 withDSM under DSM 11181 on Sep. 17, 1996.

FIG. 3 shows the base sequence (SEQ ID NOS:7 and 9) and the amino acidsequence (SEQ ID NO:8), derived therefrom, of a DNA coding for a peptideof L1 of a papilloma virus. This DNA was deposited as plasmid DL27 withDSM under DSM 11182 on Sep. 17, 1996.

FIG. 4 shows the base sequence (SEQ ID NOS:10 and 12) and the amino acidsequence (SEQ ID NO:11), derived therefrom, of a DNA coding for apeptide of L1 of a papilloma virus. This DNA was deposited as plasmidD135 with DSM under DSM 11183 on Sep. 17, 1996.

FIG. 5 shows the base sequence (SEQ ID NOS:13 and 15) and the amino acidsequence (SEQ ID NO:14), derived therefrom, of a DNA coding for apeptide of L1 of a papilloma virus. This DNA was deposited as plasmidDL40 with DSM under DSM 11184 on Sep. 17, 1996.

FIG. 6 shows the base sequence (SEQ ID NOS:16 and 18) and the amino acidsequence (SEQ ID NO:17), derived therefrom, of a DNA coding for apeptide of L1 of a papilloma virus. This DNA was deposited as plasmidDL78 with DSM under DSM 11185 on Sep. 17, 1996.

FIG. 7 shows the base sequence (SEQ ID NOS:19 and 21) and the amino acidsequence (SEQ ID NO:20), derived therefrom, of a DNA coding for apeptide of L1 of a papilloma virus. This DNA was deposited as plasmidDL82 with DSM under DSM 11186 on Sep. 17, 1996.

FIG. 8 shows the base sequence (SEQ ID NOS:22 and 24) and the amino acidsequence (SEQ ID NO:23), derived therefrom, of a DNA coding for apeptide of L1 of a papilloma virus. This DNA was deposited as plasmidDL83 with DSM under DSM 11187 on Sep. 17, 1996.

FIG. 9 shows the base sequence (SEQ ID NOS:25 and 27) and the amino acidsequence (SEQ ID NO:26), derived therefrom, of a DNA coding for apeptide of L1 of a papilloma virus. This DNA was deposited as plasmidDL84 with DSM under DSM 11188 on Sep. 17, 1996.

FIG. 10 shows the base sequence (SEQ ID NOS:28 and 30) and the aminoacid sequence (SEQ ID NO:29), derived therefrom, of a DNA coding for apeptide of L1 of papilloma virus. This DNA was deposited as plasmidDL100 with DSM under DSM 11189 on Sep. 17, 1996.

FIG. 11 shows the base sequence (SEQ ID NOS:31 and 33) and the aminoacid sequence (SEQ ID NO:32), derived therefrom, of a DNA coding for apeptide of L1 of a papilloma virus. This DNA was deposited as plasmidHR22 with DSM under DSM 11190 on Sep. 17, 1996.

V. DETAILED DESCRIPTION OF THE INVENTION

The subject matter of the invention relates to a DNA coding for apeptide of a papilloma virus major capsid protein (L1), the peptidecomprising the amino acid sequence of FIG. 1 (SEQ ID NO:2), FIG. 2 (SEQID NO:5), FIG. 3 (SEQ ID NO:8), FIG. 4 (SEQ ID NO:11), FIG. 5 (SEQ IDNO:14), FIG. 6 (SEQ ID NO:17), FIG. 7 (SEQ ID NO:20), FIG. 8 (SEQ IDNO:23), FIG. 9 (SEQ ID NO:26), FIG. 10 (SEQ ID NO:29) or FIG. 11 (SEQ IDNO:32) or an amino acid sequence differing therefrom by one or moreamino acids.

A further subject matter of the invention relates to a DNA coding for apeptide of a papilloma virus major capsid protein, the DNA comprisingthe base sequence of FIG. 1 (SEQ ID NOS:1 and 3), FIG. 2 (SEQ ID NOS:4and 6), FIG. 3 (SEQ ID NOS:7 and 9), FIG. 4 (SEQ ID NOS:10 and 12), FIG.5 (SEQ ID NOS:13 and 15), FIG. 6 (SEQ ID NOS:16 and 18), FIG. 7 (SEQ IDNOS:19 and 21), FIG. 8 (SEQ ID NOS:22 and 24), FIG. 9 (SEQ ID NOS:25 and27), FIG. 10 (SEQ ID NOS:28 and 30) or FIG. 11 (SEQ ID NOS:31 and 33) ora base sequence differing therefrom by one or more base pairs.

The above DNA was compared with the DNA of known papilloma viruses.Sequence homology studies were carried out. A homology having less than90% shows that a DNA according to the invention is a new HP virus. TheDNAs according to the invention have the following sequence homologieswith respect to known papilloma viruses:

DNA of FIG. 1: 67% with respect to HP virus 65

DNA of FIG. 2: 62% with respect to HP virus 17

DNA of FIG. 3: 78% with respect to HP virus 65

DNA of FIG. 4: |

DNA of FIG. 5: 86% with respect to HP virus 10

DNA of FIG. 6: 86% with respect to HP virus 10

DNA of FIG. 7; 62% with respect to HP virus 8

DNA of FIG. 8: 66% with respect to HP virus 65

DNA of FIG. 9: 64% with respect to HP virus 65

DNA of FIG. 10: 75% with respect to HP virus 15

DNA of FIG. 11: 81% with respect to HP virus 22 and 23, respectively.

According to the invention, the above DNA can be present in a vector andexpression vector, respectively. A person skilled in the art is familiarwith examples thereof. In the case of an expression vector for E. colithese are e.g. pGEMEX, pUC derivatives, pGEM-T and pGEX-2T. For theexpression in yeast e.g. pY100 and Ycpad1 have to be mentioned, whilefor the expression in animal cells e.g. pKCR, pEF-BOS, cDM8 and pCEV4have to be indicated.

The person skilled in the arts knows suitable cells to express the aboveDNA present in an expression vector. Examples of such cells comprise theE. coli strains HB101, DH1, x1776, JM101, JM109, and XL1-Blue, the yeaststrain Saccharomyces cerevisiae and the animal cells L, NH-3T3, FM3A,CHO, COS, Vero, and HeLa.

The person skilled in the art knows in which way the above DNA has to beinserted in an expression vector. He is also familiar with the fact thatthe above DNA can be inserted in connection with a DNA coding foranother protein and peptide, respectively, so that the above DNA can beexpressed in the form of a fusion protein.

A further subject matter of the invention relates to a papilloma virusgenome which comprises the above DNA. The expression “papilloma virusgenome” also comprises an incomplete genome, i.e. fragments of apapilloma virus genome, which comprise the above DNA. This may be e.g. aDNA coding for L1 or a portion thereof.

A common process can be used for the provision of the above papillomavirus genome. It is favorable to use a process which comprises thefollowing processing steps:

(a) isolation of the total DNA from a biopsy of epithelial neoplasm,

(b) hybridization of the total DNA of (a) with the above DNA so as todetect a papilloma virus genome included in the total DNA of (a), and

(c) cloning of the total DNA of (a) containing the papilloma virusgenome, in a vector and optionally subcloning the resulting clone, allprocessing steps originating from common DNA recombination technique.

As far as the isolation, hybridization and cloning of cell DNA isconcerned, reference is made by way of supplement to Sambrook et al.,Molecular Cloning, A Laboratory Manual, second edition, Cold SpringHarbor Laboratory (1989).

The expression “epithelial neoplasm” comprises any neoplasms ofepithelium in man and animal. Examples of such neoplasms are warts,condylomas in the genital zone and carcinomas of the skin. The latterare used preferably to isolate the above papilloma virus genome.

The expression “vector” comprises any vectors suitable for cloningchromosomal DNA and extra chromosomal DNA, respectively. Examples ofsuch vectors are cosmids such as pWE15 and Super Cos1, and phages suchas λ-phages, e.g. λZAP expressvector, λZAPII vector and λgt10 vector. Inthe present case, λ-phages are used preferably. The above vectors areknown and obtainable from the company of Stratagene.

Papilloma virus genomes according to the invention may be present inintegrated form in chromosomal DNA or in extra chromosomal fashion. Theperson skilled in the art is familiar with processes serving theclarification thereof. He also knows processes serving for finding outthe optimum restriction enzymes for cloning the papilloma virus genomes.He will orient himself by genomes of known papilloma viruses. Inparticular, the person skilled in the art will pay correspondingattention to the above-mentioned HP viruses.

The provision of a papilloma virus genome referred to as DL17-G isdescribed by way of example. For this purpose, the total DNA is isolatedfrom a biopsy of a squamous cell carcinoma, cleaved by BamHI andseparated electrophoretically in an agarose gel. The agarose gel is thensubjected to a blotting method so as to transfer the DNA to anitrocellulose membrane. It is inserted in a hybridization method inwhich the DNA of FIG. 1 is used as labeled sample, optionally incombination with a DNA of HP virus 65. Hybridization with the papillomavirus DNA present in the total DNA is obtained.

Moreover, the above total DNA cleaved by BamHI is cloned in a λ-phage.The corresponding clones, i.e. the clones containing the papilloma virusDNA are identified by hybridization with the DNA of FIG. 1, optionallyin combination with a DNA of the HP virus 65. The insert of these clonesis then subjected to a further cloning in a plasmid vector so as toobtain a clone which contains the papilloma virus genome DL17-G. Thegenome is confirmed by sequencing.

Further papilloma virus genomes are provided analogously. They aredesignated in accordance with the DNAs used for their provision, namelyby: DL20-G, DL27-G, DL35-G, DL40-G, DL78-G, DL82-G, DL-83G, DL84-G,DL100-G and HR22-G, respectively.

A further subject matter of the invention relates to a protein which iscoded by the above papilloma virus genome. Such a protein is e.g. amajor capsid protein (L1) or a minor capsid protein (L2). An aboveprotein is prepared as usual. The preparation of L1 and L2,respectively, of the papilloma virus genome DL17-G is described by wayof example. For this purpose, the HP virus 65 related to the DNA of FIG.1 (SEQ ID NOS:1 and 3) is used. The full sequence and the position ofindividual DNA regions coding for proteins are known in connectiontherewith. These DNAs are identified on the papilloma virus genomeDL17-G by parallel restriction cleavages of both genomes and subsequenthybridization with various fragments concerning the DNA encoding L1 andL2, respectively. They are confirmed by sequencing. The DNA coding forL1 is referred to as DL17-G-L1 DNA and the DNA coding for L2 is referredto as DL17-G-L2 DNA.

Furthermore, the DNA coding for L1 and L2, respectively, is inserted inan expression vector. Examples thereof are mentioned above for E. coli,yeast and animal cells. In particular, reference is made to the vectorpGEX-2T as regards the expression in E. coli (Kirnbauer et al., supra).Having inserted the DL17-G-L1 DNA and DL17-G-L2 DNA, one obtainspGEX-2T-DL17-G-L1 and pGEX-2T-DL17-G-L2, respectively. Aftertransforming E. coli, these expression vectors express a glutathione Stransferase L1 fusion protein and glutathione S transferase L2 fusionprotein, respectively. The proteins are purified as usual.

The bacculovirus system and vaccinia virus system, respectively, ismentioned for a further expression of the above DNA encoding L1 and L2,respectively. Expression vectors usable for this purpose are e.g. pEVmod. and pSynwtVI⁻ for the bacculovirus system (Kirnbauer et al.,supra). Especially vectors with the vaccinia virus “early” (p7.5k)promoter and “late” (Psynth, p11K) promoter, respectively, have to bementioned for the vaccinia virus system (Hagensee et al., 1993, Journalof Virology 67:315-322). The bacculovirus system is preferred in thepresent case. Having inserted the above DNA encoding L1 and L2,respectively, in pEV mod., one obtains pEVmod.-DL17-G-L1 andpEVmod.-DL17-C-L2, respectively.

The former expression vector as such or both expression vectors jointlylead to the formation of virus-like particles after infection of SF-9insect cells. In the former case, such a particle comprises an L1protein, while in the latter case it contains an L2 protein in additionto an L1 protein.

A virus-like particle of the latter case is also obtained by insertingthe above DL17-G-L1 and DL17-G-L2 DNAs jointly in the expression vectorpSynwtVI⁻ and using the resulting pSynwtVI⁻DL17-G-L1/L2 for theinfection of SF-9 insect cells. The above virus-like particles arepurified as usual. They also represent a subject matter of theinvention.

A further subject matter of the invention relates to an antibodydirected against an above protein and virus-like particle, respectively.The preparation thereof is made as usual. It is described by way ofexample for the preparation of an antibody which is directed against avirus-like particle comprising L1 of DL17-G. For this purpose, thevirus-like particle is injected subcutaneously into BALB/c mice. Thisinjection is repeated at intervals of 3 weeks each. About 2 weeks afterthe last injection, the serum containing the antibody is isolated andtested as usual.

In a preferred embodiment, the antibody is a monoclonal antibody. Forits preparation, spleen cells are removed from the mice after the abovefourth injection and fused with myeloma cells as usual. The furthercloning also takes place according to known methods.

By means of the present invention it is possible to detect papillomaviruses, particularly in carcinomas of the skin. For this purpose, theDNA according to the invention can be used as such or when comprised bya further DNA. The latter may also be a papilloma virus genome or aportion thereof.

The present invention also enables the provision of formerly unknownpapilloma viruses. They are found especially in carcinomas of the skin.In addition, the invention supplies proteins and virus-like particleswhich originate from these papilloma viruses. Moreover, antibodies areprovided which are directed against these proteins and particles,respectively.

The present invention also enables to take diagnostic and therapeuticsteps in the case of papilloma virus diseases. Moreover, it supplies thepossibility of building up a vaccine against papilloma virus infections.Thus, the present invention represents a break-through in the field ofpapilloma virus research.

The below examples explain the invention in more detail. The followingpreparations and examples are given to enable those skilled in the artto more clearly understand and to practice the present invention. Thepresent invention, however, is not limited in scope by the exemplifiedembodiments, which are intended as illustrations of single aspects ofthe invention only, and methods which are functionally equivalent arewithin the scope of the invention. Indeed, various modifications of theinvention in addition to those described herein will become apparent tothose skilled in the art from the foregoing description and accompanyingdrawings. Such modifications are intended to fall within the scope ofthe appended claims.

VI. EXAMPLES A. Example 1 Identification of the Papilloma Virus GenomeDL17-G

The total DNA is isolated from a biopsy of a squamous cell carcinoma. 10μg of this DNA are cleaved by the restriction enzyme BamHI and separatedelectrophoretically in a 0.5% agarose gel. At the same time, 10 μg ofthe above DNA which was not cleaved, is also separated. The agarose gelis subjected to a blotting method so as to transfer the DNA from theagarose gel to a nitrocellulose membrane. It is employed in ahybridization method in which the above DNA of FIG. 1 (SEQ ID NOS:1 and3) is used in combination with the HP virus-65 DNA as p³²-labeledsample. Hybridization with the blotted DNA is obtained.

The person skilled in the field of DNA recombination technique isfamiliar with the above methods. Reference is made to Sambrook et al.,supra, by way of supplement.

B. Example 2 Cloning of the Papilloma Virus Genome DL17-G

The biopsy DNA obtained from Example 1 is cleaved by the restrictionenzyme BamHI. The resulting fragments are used in a ligase reaction inwhich the dephosphorylated vector λZAP express cleaved by BamHI is alsopresent. The resulting recombinant DNA molecules are packed inbacteriophages, and they are used for infecting bacteria. For theseprocessing steps, the ZAP express vector kit offered by the company ofStratagene is used. The resulting phage plaques are then subjected to ahybridization process which uses the p³²-labeled DNA of FIG. 1 (SEQ IDNOS:1 and 3) employed in Example 1 in combination with p³²-labeled HPvirus-65 DNA. Hybridization with corresponding phage plaques isobtained. The BamHI fragments of DL17-G are isolated therefrom and usedin a further ligase reaction together with a BamHI-cleaved,dephosphorylated plasmid vector, pBluescript. The resulting recombinantDNA molecules are used for transforming bacteria, E. coli XL1-Blue. Byrestriction cleavages and hybridization with the above DNA samples,respectively, a bacterial clone containing the papilloma virus genomeDL17-G is identified. The plasmid of this bacterial clone is referred toas pBlue-DL17-G.

All references cited within the body of the instant specification arehereby incorporated by reference in their entirety.

33 1 416 DNA Papilloma virus CDS (1)...(414) 1 ctc gag gat ggg gaa atgtgc gat att gga ttt gga gca tgc aat ttt 48 Leu Glu Asp Gly Glu Met CysAsp Ile Gly Phe Gly Ala Cys Asn Phe 1 5 10 15 aaa cag tta cag aag gataga tct ggt gtt cca tta gat ata gta gag 96 Lys Gln Leu Gln Lys Asp ArgSer Gly Val Pro Leu Asp Ile Val Glu 20 25 30 agc act tgt aag tat cct gatttt tta aag atg ggt aag gat atg tat 144 Ser Thr Cys Lys Tyr Pro Asp PheLeu Lys Met Gly Lys Asp Met Tyr 35 40 45 ggc gac gag cta ttt ttc tat ggcaga cga gaa cag tta tat gta aga 192 Gly Asp Glu Leu Phe Phe Tyr Gly ArgArg Glu Gln Leu Tyr Val Arg 50 55 60 cat aac ttt tct cgt gca ggc act gtagga gat agt ata cct tta cct 240 His Asn Phe Ser Arg Ala Gly Thr Val GlyAsp Ser Ile Pro Leu Pro 65 70 75 80 gat cag gat act gca ttt tat aga agtcct aat act gca aat aat gat 288 Asp Gln Asp Thr Ala Phe Tyr Arg Ser ProAsn Thr Ala Asn Asn Asp 85 90 95 ttg cct caa aat aca tta gcg tct cac atatac tgt gcc atc cca agt 336 Leu Pro Gln Asn Thr Leu Ala Ser His Ile TyrCys Ala Ile Pro Ser 100 105 110 ggt tct tta acc agt agt gat tct caa ttattt aat agg cct tat tgg 384 Gly Ser Leu Thr Ser Ser Asp Ser Gln Leu PheAsn Arg Pro Tyr Trp 115 120 125 ctg caa aat gct cag ggc acc aac aac ggcgt 416 Leu Gln Asn Ala Gln Gly Thr Asn Asn Gly 130 135 2 138 PRTPapilloma virus 2 Leu Glu Asp Gly Glu Met Cys Asp Ile Gly Phe Gly AlaCys Asn Phe 1 5 10 15 Lys Gln Leu Gln Lys Asp Arg Ser Gly Val Pro LeuAsp Ile Val Glu 20 25 30 Ser Thr Cys Lys Tyr Pro Asp Phe Leu Lys Met GlyLys Asp Met Tyr 35 40 45 Gly Asp Glu Leu Phe Phe Tyr Gly Arg Arg Glu GlnLeu Tyr Val Arg 50 55 60 His Asn Phe Ser Arg Ala Gly Thr Val Gly Asp SerIle Pro Leu Pro 65 70 75 80 Asp Gln Asp Thr Ala Phe Tyr Arg Ser Pro AsnThr Ala Asn Asn Asp 85 90 95 Leu Pro Gln Asn Thr Leu Ala Ser His Ile TyrCys Ala Ile Pro Ser 100 105 110 Gly Ser Leu Thr Ser Ser Asp Ser Gln LeuPhe Asn Arg Pro Tyr Trp 115 120 125 Leu Gln Asn Ala Gln Gly Thr Asn AsnGly 130 135 3 416 DNA Papilloma virus 3 acgccgttgt tggtgccctg agcattttgcagccaataag gcctattaaa taattgagaa 60 tcactactgg ttaaagaacc acttgggatggcacagtata tgtgagacgc taatgtattt 120 tgaggcaaat cattatttgc agtattaggacttctataaa atgcagtatc ctgatcaggt 180 aaaggtatac tatctcctac agtgcctgcacgagaaaagt tatgtcttac atataactgt 240 tctcgtctgc catagaaaaa tagctcgtcgccatacatat ccttacccat ctttaaaaaa 300 tcaggatact tacaagtgct ctctactatatctaatggaa caccagatct atccttctgt 360 aactgtttaa aattgcatgc tccaaatccaatatcgcaca tttccccatc ctcgag 416 4 386 DNA Papilloma virus CDS(1)...(384) 4 att gag gat ggt gat atg gta gac ata gga ttt ggg aat tgtaat ttt 48 Ile Glu Asp Gly Asp Met Val Asp Ile Gly Phe Gly Asn Cys AsnPhe 1 5 10 15 aaa gct tta caa cag gac aag gct ggt acg cct tta gag ttaact aat 96 Lys Ala Leu Gln Gln Asp Lys Ala Gly Thr Pro Leu Glu Leu ThrAsn 20 25 30 gaa aaa tgt aag tgg cca gat ttt ctg aag atg gaa aaa gac acttat 144 Glu Lys Cys Lys Trp Pro Asp Phe Leu Lys Met Glu Lys Asp Thr Tyr35 40 45 gga gac cag atg ttt ttc tgt ggc agg aag gag caa atg tat tct agg192 Gly Asp Gln Met Phe Phe Cys Gly Arg Lys Glu Gln Met Tyr Ser Arg 5055 60 cat atg ctt gct aaa gcg ggt att gat ggt gat cat ata cca gaa tca240 His Met Leu Ala Lys Ala Gly Ile Asp Gly Asp His Ile Pro Glu Ser 6570 75 80 tta tac cat tcg cca aag aat aat gga aat ggc att gct cct tac act288 Leu Tyr His Ser Pro Lys Asn Asn Gly Asn Gly Ile Ala Pro Tyr Thr 8590 95 tac ttt cca acc aca agc ggt tcc tta gtc aca agt gat aat caa tta336 Tyr Phe Pro Thr Thr Ser Gly Ser Leu Val Thr Ser Asp Asn Gln Leu 100105 110 ttt aac agg cca tat tgg ctt cat aat tca caa gga acc aat aac ggt384 Phe Asn Arg Pro Tyr Trp Leu His Asn Ser Gln Gly Thr Asn Asn Gly 115120 125 at 386 5 128 PRT Papilloma virus 5 Ile Glu Asp Gly Asp Met ValAsp Ile Gly Phe Gly Asn Cys Asn Phe 1 5 10 15 Lys Ala Leu Gln Gln AspLys Ala Gly Thr Pro Leu Glu Leu Thr Asn 20 25 30 Glu Lys Cys Lys Trp ProAsp Phe Leu Lys Met Glu Lys Asp Thr Tyr 35 40 45 Gly Asp Gln Met Phe PheCys Gly Arg Lys Glu Gln Met Tyr Ser Arg 50 55 60 His Met Leu Ala Lys AlaGly Ile Asp Gly Asp His Ile Pro Glu Ser 65 70 75 80 Leu Tyr His Ser ProLys Asn Asn Gly Asn Gly Ile Ala Pro Tyr Thr 85 90 95 Tyr Phe Pro Thr ThrSer Gly Ser Leu Val Thr Ser Asp Asn Gln Leu 100 105 110 Phe Asn Arg ProTyr Trp Leu His Asn Ser Gln Gly Thr Asn Asn Gly 115 120 125 6 386 DNAPapilloma virus 6 ataccgttat tggttccttg tgaattatga agccaatatg gcctgttaaataattgatta 60 tcacttgtga ctaaggaacc gcttgtggtt ggaaagtaag tgtaaggagcaatgccattt 120 ccattattct ttggcgaatg gtataatgat tctggtatat gatcaccatcaatacccgct 180 ttagcaagca tatgcctaga atacatttgc tccttcctgc cacagaaaaacatctggtct 240 ccataagtgt ctttttccat cttcagaaaa tctggccact tacatttttcattagttaac 300 tctaaaggcg taccagcctt gtcctgttgt aaagctttaa aattacaattcccaaatcct 360 atgtctacca tatcaccatc ctcaat 386 7 410 DNA Papillomavirus CDS (1)...(408) 7 ctg gag gat ggc gac atg tgt gat ata ggc ttt ggagct ttt aac ttt 48 Leu Glu Asp Gly Asp Met Cys Asp Ile Gly Phe Gly AlaPhe Asn Phe 1 5 10 15 aaa gct ttg cag gat gat aaa tcc agt gca cca ttagat gta gtt ggt 96 Lys Ala Leu Gln Asp Asp Lys Ser Ser Ala Pro Leu AspVal Val Gly 20 25 30 act ttg tgt aaa tgg cct gac ttc tta aag atg agt aaggac att tat 144 Thr Leu Cys Lys Trp Pro Asp Phe Leu Lys Met Ser Lys AspIle Tyr 35 40 45 ggt gac agt tta ttc ttc ttt ggc cga agg gaa cag ctt tatgca aga 192 Gly Asp Ser Leu Phe Phe Phe Gly Arg Arg Glu Gln Leu Tyr AlaArg 50 55 60 cac ttt ttt gtt aga gct ggg aca atg ggt gat gcg tta cca gagcct 240 His Phe Phe Val Arg Ala Gly Thr Met Gly Asp Ala Leu Pro Glu Pro65 70 75 80 ttt gaa gtg aag tct gat tac ata att gct gct cag agt aac caagaa 288 Phe Glu Val Lys Ser Asp Tyr Ile Ile Ala Ala Gln Ser Asn Gln Glu85 90 95 caa aat aat ctt ggc cct cac att tat ttt gga act cct agc ggt tct336 Gln Asn Asn Leu Gly Pro His Ile Tyr Phe Gly Thr Pro Ser Gly Ser 100105 110 ctt gta tca agt gaa tct cag ctt ttt aac cga ccg tat tgg tta aac384 Leu Val Ser Ser Glu Ser Gln Leu Phe Asn Arg Pro Tyr Trp Leu Asn 115120 125 aga gct cag ggc act aat aac ggc at 410 Arg Ala Gln Gly Thr AsnAsn Gly 130 135 8 136 PRT Papilloma virus 8 Leu Glu Asp Gly Asp Met CysAsp Ile Gly Phe Gly Ala Phe Asn Phe 1 5 10 15 Lys Ala Leu Gln Asp AspLys Ser Ser Ala Pro Leu Asp Val Val Gly 20 25 30 Thr Leu Cys Lys Trp ProAsp Phe Leu Lys Met Ser Lys Asp Ile Tyr 35 40 45 Gly Asp Ser Leu Phe PhePhe Gly Arg Arg Glu Gln Leu Tyr Ala Arg 50 55 60 His Phe Phe Val Arg AlaGly Thr Met Gly Asp Ala Leu Pro Glu Pro 65 70 75 80 Phe Glu Val Lys SerAsp Tyr Ile Ile Ala Ala Gln Ser Asn Gln Glu 85 90 95 Gln Asn Asn Leu GlyPro His Ile Tyr Phe Gly Thr Pro Ser Gly Ser 100 105 110 Leu Val Ser SerGlu Ser Gln Leu Phe Asn Arg Pro Tyr Trp Leu Asn 115 120 125 Arg Ala GlnGly Thr Asn Asn Gly 130 135 9 410 DNA Papilloma virus 9 atgccgttattagtgccctg agctctgttt aaccaatacg gtcggttaaa aagctgagat 60 tcacttgatacaagagaacc gctaggagtt ccaaaataaa tgtgagggcc aagattattt 120 tgttcttggttactctgagc agcaattatg taatcagact tcacttcaaa aggctctggt 180 aacgcatcacccattgtccc agctctaaca aaaaagtgtc ttgcataaag ctgttccctt 240 cggccaaagaagaataaact gtcaccataa atgtccttac tcatctttaa gaagtcaggc 300 catttacacaaagtaccaac tacatctaat ggtgcactgg atttatcatc ctgcaaagct 360 ttaaagttaaaagctccaaa gcctatatca cacatgtcgc catcctccag 410 10 383 DNA Papillomavirus CDS (1)...(381) 10 ata gag gat ggg gaa atg gtt gaa act ggg ttc ggggcc ctg gat ttt 48 Ile Glu Asp Gly Glu Met Val Glu Thr Gly Phe Gly AlaLeu Asp Phe 1 5 10 15 gcc gct cta cag tcc aac aaa tct gat gtc ccc ctggat att tgt act 96 Ala Ala Leu Gln Ser Asn Lys Ser Asp Val Pro Leu AspIle Cys Thr 20 25 30 aac ata tgt aaa tat ccg gac tat ctg aaa atg gct gctgac ccc tat 144 Asn Ile Cys Lys Tyr Pro Asp Tyr Leu Lys Met Ala Ala AspPro Tyr 35 40 45 ggc gat tct atg ttc ttt tcc ctg cgc agg gag cag atg ttcacc cgg 192 Gly Asp Ser Met Phe Phe Ser Leu Arg Arg Glu Gln Met Phe ThrArg 50 55 60 cat ttc ttc aat cgg ggt ggg tcg atg ggt gac gcc ctc ccg gaggag 240 His Phe Phe Asn Arg Gly Gly Ser Met Gly Asp Ala Leu Pro Glu Glu65 70 75 80 cta tac gtc aaa agt tct acc gtg cag acc cca ggt agt tat gtttac 288 Leu Tyr Val Lys Ser Ser Thr Val Gln Thr Pro Gly Ser Tyr Val Tyr85 90 95 acc tcc act ccc agt ggc tct atg gta tcc tct gaa cag cag tta ttt336 Thr Ser Thr Pro Ser Gly Ser Met Val Ser Ser Glu Gln Gln Leu Phe 100105 110 aac aag cct tac tgg ctg cgg agg gct caa ggt act aat aac ggc 381Asn Lys Pro Tyr Trp Leu Arg Arg Ala Gln Gly Thr Asn Asn Gly 115 120 125gt 383 11 127 PRT Papilloma virus 11 Ile Glu Asp Gly Glu Met Val Glu ThrGly Phe Gly Ala Leu Asp Phe 1 5 10 15 Ala Ala Leu Gln Ser Asn Lys SerAsp Val Pro Leu Asp Ile Cys Thr 20 25 30 Asn Ile Cys Lys Tyr Pro Asp TyrLeu Lys Met Ala Ala Asp Pro Tyr 35 40 45 Gly Asp Ser Met Phe Phe Ser LeuArg Arg Glu Gln Met Phe Thr Arg 50 55 60 His Phe Phe Asn Arg Gly Gly SerMet Gly Asp Ala Leu Pro Glu Glu 65 70 75 80 Leu Tyr Val Lys Ser Ser ThrVal Gln Thr Pro Gly Ser Tyr Val Tyr 85 90 95 Thr Ser Thr Pro Ser Gly SerMet Val Ser Ser Glu Gln Gln Leu Phe 100 105 110 Asn Lys Pro Tyr Trp LeuArg Arg Ala Gln Gly Thr Asn Asn Gly 115 120 125 12 383 DNA Papillomavirus 12 acgccgttat tagtaccttg agccctccgc agccagtaag gcttgttaaataactgctgt 60 tcagaggata ccatagagcc actgggagtg gaggtgtaaa cataactacctggggtctgc 120 acggtagaac ttttgacgta tagctcctcc gggagggcgt cacccatcgacccaccccga 180 ttgaagaaat gccgggtgaa catctgctcc ctgcgcaggg aaaagaacatagaatcgcca 240 taggggtcag cagccatttt cagatagtcc ggatatttac atatgttagtacaaatatcc 300 agggggacat cagatttgtt ggactgtaga gcggcaaaat ccagggccccgaacccagtt 360 tcaaccattt ccccatcctc tat 383 13 386 DNA Papilloma virusCDS (1)...(384) 13 atg gaa gac gga gat atg gta gac act ggc tat ggt gctatg gac ttc 48 Met Glu Asp Gly Asp Met Val Asp Thr Gly Tyr Gly Ala MetAsp Phe 1 5 10 15 act gca tta cag tta aat aag tct gac gtg cct ata gatatt tgc cag 96 Thr Ala Leu Gln Leu Asn Lys Ser Asp Val Pro Ile Asp IleCys Gln 20 25 30 tcc act tgt aaa tac cct gat tat ttg ggc atg gca gca gagcct tat 144 Ser Thr Cys Lys Tyr Pro Asp Tyr Leu Gly Met Ala Ala Glu ProTyr 35 40 45 ggc gac agc atg ttt ttt tat ttg cgc aga gag caa ctg ttt gcaaga 192 Gly Asp Ser Met Phe Phe Tyr Leu Arg Arg Glu Gln Leu Phe Ala Arg50 55 60 cat ttt ttc aat aga gcc agt gca gtg gga gac acc att cct gac act240 His Phe Phe Asn Arg Ala Ser Ala Val Gly Asp Thr Ile Pro Asp Thr 6570 75 80 tta ata ttg aag tcg gcc agt ggt gac caa aac gtt ggt agt gct gtg288 Leu Ile Leu Lys Ser Ala Ser Gly Asp Gln Asn Val Gly Ser Ala Val 8590 95 tat agc ccc act ccc agt ggg tcc atg gta aca tct gag gct caa tta336 Tyr Ser Pro Thr Pro Ser Gly Ser Met Val Thr Ser Glu Ala Gln Leu 100105 110 ttt aat aag cca tat tgg ctg aag cgg gct caa gga cat aac aat ggt384 Phe Asn Lys Pro Tyr Trp Leu Lys Arg Ala Gln Gly His Asn Asn Gly 115120 125 gt 386 14 128 PRT Papilloma virus 14 Met Glu Asp Gly Asp Met ValAsp Thr Gly Tyr Gly Ala Met Asp Phe 1 5 10 15 Thr Ala Leu Gln Leu AsnLys Ser Asp Val Pro Ile Asp Ile Cys Gln 20 25 30 Ser Thr Cys Lys Tyr ProAsp Tyr Leu Gly Met Ala Ala Glu Pro Tyr 35 40 45 Gly Asp Ser Met Phe PheTyr Leu Arg Arg Glu Gln Leu Phe Ala Arg 50 55 60 His Phe Phe Asn Arg AlaSer Ala Val Gly Asp Thr Ile Pro Asp Thr 65 70 75 80 Leu Ile Leu Lys SerAla Ser Gly Asp Gln Asn Val Gly Ser Ala Val 85 90 95 Tyr Ser Pro Thr ProSer Gly Ser Met Val Thr Ser Glu Ala Gln Leu 100 105 110 Phe Asn Lys ProTyr Trp Leu Lys Arg Ala Gln Gly His Asn Asn Gly 115 120 125 15 386 DNAPapilloma virus 15 acaccattgt tatgtccttg agcccgcttc agccaatatggcttattaaa taattgagcc 60 tcagatgtta ccatggaccc actgggagtg gggctatacacagcactacc aacgttttgg 120 tcaccactgg ccgacttcaa tattaaagtg tcaggaatggtgtctcccac tgcactggct 180 ctattgaaaa aatgtcttgc aaacagttgc tctctgcgcaaataaaaaaa catgctgtcg 240 ccataaggct ctgctgccat gcccaaataa tcagggtatttacaagtgga ctggcaaata 300 tctataggca cgtcagactt atttaactgt aatgcagtgaagtccatagc accatagcca 360 gtgtctacca tatctccgtc ttccat 386 16 386 DNAPapilloma virus CDS (1)...(384) 16 ctg gag gat gcg gaa atg gta gac actgga tat ggt gcc atg gac ttt 48 Leu Glu Asp Ala Glu Met Val Asp Thr GlyTyr Gly Ala Met Asp Phe 1 5 10 15 act gca tta cag tta aat aag tct gacgtg cct atc gat att tgc cag 96 Thr Ala Leu Gln Leu Asn Lys Ser Asp ValPro Ile Asp Ile Cys Gln 20 25 30 tct acc tgt aaa tat cct gat tat ttg ggcatg gca gca gag cct tat 144 Ser Thr Cys Lys Tyr Pro Asp Tyr Leu Gly MetAla Ala Glu Pro Tyr 35 40 45 ggc gac agc atg ttt ttt tat ttg cgc agg gaacaa ctg ttt gcc aga 192 Gly Asp Ser Met Phe Phe Tyr Leu Arg Arg Glu GlnLeu Phe Ala Arg 50 55 60 cat ttt ttt aat agg gct agt gca gtt ggg gac accatt cct gac act 240 His Phe Phe Asn Arg Ala Ser Ala Val Gly Asp Thr IlePro Asp Thr 65 70 75 80 ttg ata ttg aag gca gcc agt gga ggg caa aac gttggt agt gct gtt 288 Leu Ile Leu Lys Ala Ala Ser Gly Gly Gln Asn Val GlySer Ala Val 85 90 95 tac agc ccc aca ccc agt ggg tcc atg gta aca tct gaggct caa ttg 336 Tyr Ser Pro Thr Pro Ser Gly Ser Met Val Thr Ser Glu AlaGln Leu 100 105 110 ttt aat aag cca tat tgg cta cgg cgg gct caa gga acgaac aac gga 384 Phe Asn Lys Pro Tyr Trp Leu Arg Arg Ala Gln Gly Thr AsnAsn Gly 115 120 125 gt 386 17 128 PRT Papilloma virus 17 Leu Glu Asp AlaGlu Met Val Asp Thr Gly Tyr Gly Ala Met Asp Phe 1 5 10 15 Thr Ala LeuGln Leu Asn Lys Ser Asp Val Pro Ile Asp Ile Cys Gln 20 25 30 Ser Thr CysLys Tyr Pro Asp Tyr Leu Gly Met Ala Ala Glu Pro Tyr 35 40 45 Gly Asp SerMet Phe Phe Tyr Leu Arg Arg Glu Gln Leu Phe Ala Arg 50 55 60 His Phe PheAsn Arg Ala Ser Ala Val Gly Asp Thr Ile Pro Asp Thr 65 70 75 80 Leu IleLeu Lys Ala Ala Ser Gly Gly Gln Asn Val Gly Ser Ala Val 85 90 95 Tyr SerPro Thr Pro Ser Gly Ser Met Val Thr Ser Glu Ala Gln Leu 100 105 110 PheAsn Lys Pro Tyr Trp Leu Arg Arg Ala Gln Gly Thr Asn Asn Gly 115 120 12518 386 DNA Papilloma virus 18 actccgttgt tcgttccttg agcccgccgtagccaatatg gcttattaaa caattgagcc 60 tcagatgtta ccatggaccc actgggtgtggggctgtaaa cagcactacc aacgttttgc 120 cctccactgg ctgccttcaa tatcaaagtgtcaggaatgg tgtccccaac tgcactagcc 180 ctattaaaaa aatgtctggc aaacagttgttccctgcgca aataaaaaaa catgctgtcg 240 ccataaggct ctgctgccat gcccaaataatcaggatatt tacaggtaga ctggcaaata 300 tcgataggca cgtcagactt atttaactgtaatgcagtaa agtccatggc accatatcca 360 gtgtctacca tttccgcatc ctccag 386 19413 DNA Papilloma virus CDS (1)...(411) 19 atc gag gat ggg gat atg tgtgat att ggt ttt gga aac atg aat ttc 48 Ile Glu Asp Gly Asp Met Cys AspIle Gly Phe Gly Asn Met Asn Phe 1 5 10 15 agt ata tta caa caa gac agatca ggt gtt cct ttg gat ata gta gct 96 Ser Ile Leu Gln Gln Asp Arg SerGly Val Pro Leu Asp Ile Val Ala 20 25 30 tcc att tgc aaa tgg cca gat cttggt aaa atg acc aat gat gtg tat 144 Ser Ile Cys Lys Trp Pro Asp Leu GlyLys Met Thr Asn Asp Val Tyr 35 40 45 ggt gat gaa cta ttc ttt ttt ggt aaacgg gag cag gtt tat gca agg 192 Gly Asp Glu Leu Phe Phe Phe Gly Lys ArgGlu Gln Val Tyr Ala Arg 50 55 60 cat tat ttt aca agg cat ggt gtt gta ggagaa gat att cct cag gta 240 His Tyr Phe Thr Arg His Gly Val Val Gly GluAsp Ile Pro Gln Val 65 70 75 80 aat gag gac cct aca act aaa tac ctg cgagga ggt gaa ggt ggt caa 288 Asn Glu Asp Pro Thr Thr Lys Tyr Leu Arg GlyGly Glu Gly Gly Gln 85 90 95 aat cag gct act gtt tca tcc tct gta tat tttgca act ccc agt ggt 336 Asn Gln Ala Thr Val Ser Ser Ser Val Tyr Phe AlaThr Pro Ser Gly 100 105 110 tcc ttg gtg tcc agt gat gct caa att atg aacagg cct tat tgg gta 384 Ser Leu Val Ser Ser Asp Ala Gln Ile Met Asn ArgPro Tyr Trp Val 115 120 125 caa cgc gcg cag gga acg aac aac ggc gt 413Gln Arg Ala Gln Gly Thr Asn Asn Gly 130 135 20 137 PRT Papilloma virus20 Ile Glu Asp Gly Asp Met Cys Asp Ile Gly Phe Gly Asn Met Asn Phe 1 510 15 Ser Ile Leu Gln Gln Asp Arg Ser Gly Val Pro Leu Asp Ile Val Ala 2025 30 Ser Ile Cys Lys Trp Pro Asp Leu Gly Lys Met Thr Asn Asp Val Tyr 3540 45 Gly Asp Glu Leu Phe Phe Phe Gly Lys Arg Glu Gln Val Tyr Ala Arg 5055 60 His Tyr Phe Thr Arg His Gly Val Val Gly Glu Asp Ile Pro Gln Val 6570 75 80 Asn Glu Asp Pro Thr Thr Lys Tyr Leu Arg Gly Gly Glu Gly Gly Gln85 90 95 Asn Gln Ala Thr Val Ser Ser Ser Val Tyr Phe Ala Thr Pro Ser Gly100 105 110 Ser Leu Val Ser Ser Asp Ala Gln Ile Met Asn Arg Pro Tyr TrpVal 115 120 125 Gln Arg Ala Gln Gly Thr Asn Asn Gly 130 135 21 413 DNAPapilloma virus 21 acgccgttgt tcgttccctg cgcgcgttgt acccaataaggcctgttcat aatttgagca 60 tcactggaca ccaaggaacc actgggagtt gcaaaatatacagaggatga aacagtagcc 120 tgattttgac caccttcacc tcctcgcagg tatttagttgtagggtcctc atttacctga 180 ggaatatctt ctcctacaac accatgcctt gtaaaataatgccttgcata aacctgctcc 240 cgtttaccaa aaaagaatag ttcatcacca tacacatcattggtcatttt accaagatct 300 ggccatttgc aaatggaagc tactatatcc aaaggaacacctgatctgtc ttgttgtaat 360 atactgaaat tcatgtttcc aaaaccaata tcacacatatccccatcctc gat 413 22 395 DNA Papilloma virus CDS (1)...(393) 22 ctg gaggat ggc gac atg tgt gat gta gga ttt gga gct gca aat ttt 48 Leu Glu AspGly Asp Met Cys Asp Val Gly Phe Gly Ala Ala Asn Phe 1 5 10 15 aaa acactt cag gaa gat aaa tca gga gta cct atg gat ctt tta aat 96 Lys Thr LeuGln Glu Asp Lys Ser Gly Val Pro Met Asp Leu Leu Asn 20 25 30 gaa act tgtaaa tat cct gac ttt ctt cag atg tca aaa gac aaa tat 144 Glu Thr Cys LysTyr Pro Asp Phe Leu Gln Met Ser Lys Asp Lys Tyr 35 40 45 gga gac agt ttgttc ttt ttt gga aga aag gaa caa ctt tac gcg aga 192 Gly Asp Ser Leu PhePhe Phe Gly Arg Lys Glu Gln Leu Tyr Ala Arg 50 55 60 cac ttt tat gtt agagga ggt gtt gat ggc gat gca ttg cca tta act 240 His Phe Tyr Val Arg GlyGly Val Asp Gly Asp Ala Leu Pro Leu Thr 65 70 75 80 aac ttt att tat ggtgct cag cag gac aaa cct caa aac aat tta gga 288 Asn Phe Ile Tyr Gly AlaGln Gln Asp Lys Pro Gln Asn Asn Leu Gly 85 90 95 cca tat act tac ttt cctact cct agt ggc tct tta tac tca acc gat 336 Pro Tyr Thr Tyr Phe Pro ThrPro Ser Gly Ser Leu Tyr Ser Thr Asp 100 105 110 aat caa tta ttt aac agaccc tat tgg ctt agc cag gct cag ggc aca 384 Asn Gln Leu Phe Asn Arg ProTyr Trp Leu Ser Gln Ala Gln Gly Thr 115 120 125 aac aat ggg at 395 AsnAsn Gly 130 23 131 PRT Papilloma virus 23 Leu Glu Asp Gly Asp Met CysAsp Val Gly Phe Gly Ala Ala Asn Phe 1 5 10 15 Lys Thr Leu Gln Glu AspLys Ser Gly Val Pro Met Asp Leu Leu Asn 20 25 30 Glu Thr Cys Lys Tyr ProAsp Phe Leu Gln Met Ser Lys Asp Lys Tyr 35 40 45 Gly Asp Ser Leu Phe PhePhe Gly Arg Lys Glu Gln Leu Tyr Ala Arg 50 55 60 His Phe Tyr Val Arg GlyGly Val Asp Gly Asp Ala Leu Pro Leu Thr 65 70 75 80 Asn Phe Ile Tyr GlyAla Gln Gln Asp Lys Pro Gln Asn Asn Leu Gly 85 90 95 Pro Tyr Thr Tyr PhePro Thr Pro Ser Gly Ser Leu Tyr Ser Thr Asp 100 105 110 Asn Gln Leu PheAsn Arg Pro Tyr Trp Leu Ser Gln Ala Gln Gly Thr 115 120 125 Asn Asn Gly130 24 395 DNA Papilloma virus 24 atcccattgt ttgtgccctg agcctggctaagccaatagg gtctgttaaa taattgatta 60 tcggttgagt ataaagagcc actaggagtaggaaagtaag tatatggtcc taaattgttt 120 tgaggtttgt cctgctgagc accataaataaagttagtta atggcaatgc atcgccatca 180 acacctcctc taacataaaa gtgtctcgcgtaaagttgtt cctttcttcc aaaaaagaac 240 aaactgtctc catatttgtc ttttgacatctgaagaaagt caggatattt acaagtttca 300 tttaaaagat ccataggtac tcctgatttatcttcctgaa gtgttttaaa atttgcagct 360 ccaaatccta catcacacat gtcgccatcctccag 395 25 407 DNA Papilloma virus CDS (1)...(405) 25 cta gaa gac gcggag atg agt gat att ggt ttg ggg gca gtt aat ttt 48 Leu Glu Asp Ala GluMet Ser Asp Ile Gly Leu Gly Ala Val Asn Phe 1 5 10 15 cat acg ttt tcggcc tcc cgt tca gat gct cct tta gaa gtt ata gat 96 His Thr Phe Ser AlaSer Arg Ser Asp Ala Pro Leu Glu Val Ile Asp 20 25 30 tca att tgc aaa tggcct gat ttt gtt cag atg aca aaa gat gtt tat 144 Ser Ile Cys Lys Trp ProAsp Phe Val Gln Met Thr Lys Asp Val Tyr 35 40 45 gga gat aag atc tgg ttttat gga aaa cgg gaa cag ctt tat gcc aga 192 Gly Asp Lys Ile Trp Phe TyrGly Lys Arg Glu Gln Leu Tyr Ala Arg 50 55 60 cat atg ttt gtt aag gat ggtgtg gac ggt gac agt att cca aat gag 240 His Met Phe Val Lys Asp Gly ValAsp Gly Asp Ser Ile Pro Asn Glu 65 70 75 80 ccc aca cac gct tat tat attccc cca ccc aca ggt tct gcc cag gaa 288 Pro Thr His Ala Tyr Tyr Ile ProPro Pro Thr Gly Ser Ala Gln Glu 85 90 95 act aat ttt gga aag ata agt tacttt cca gtt ccc agt gga tct ttg 336 Thr Asn Phe Gly Lys Ile Ser Tyr PhePro Val Pro Ser Gly Ser Leu 100 105 110 gtg tcc agt gag gct act ata tttaat aga cct tat tgg ttg cac aaa 384 Val Ser Ser Glu Ala Thr Ile Phe AsnArg Pro Tyr Trp Leu His Lys 115 120 125 gct caa gga aca aac aat gga at407 Ala Gln Gly Thr Asn Asn Gly 130 135 26 135 PRT Papilloma virus 26Leu Glu Asp Ala Glu Met Ser Asp Ile Gly Leu Gly Ala Val Asn Phe 1 5 1015 His Thr Phe Ser Ala Ser Arg Ser Asp Ala Pro Leu Glu Val Ile Asp 20 2530 Ser Ile Cys Lys Trp Pro Asp Phe Val Gln Met Thr Lys Asp Val Tyr 35 4045 Gly Asp Lys Ile Trp Phe Tyr Gly Lys Arg Glu Gln Leu Tyr Ala Arg 50 5560 His Met Phe Val Lys Asp Gly Val Asp Gly Asp Ser Ile Pro Asn Glu 65 7075 80 Pro Thr His Ala Tyr Tyr Ile Pro Pro Pro Thr Gly Ser Ala Gln Glu 8590 95 Thr Asn Phe Gly Lys Ile Ser Tyr Phe Pro Val Pro Ser Gly Ser Leu100 105 110 Val Ser Ser Glu Ala Thr Ile Phe Asn Arg Pro Tyr Trp Leu HisLys 115 120 125 Ala Gln Gly Thr Asn Asn Gly 130 135 27 407 DNA Papillomavirus 27 attccattgt ttgttccttg agctttgtgc aaccaataag gtctattaaatatagtagcc 60 tcactggaca ccaaagatcc actgggaact ggaaagtaac ttatctttccaaaattagtt 120 tcctgggcag aacctgtggg tgggggaata taataagcgt gtgtgggctcatttggaata 180 ctgtcaccgt ccacaccatc cttaacaaac atatgtctgg cataaagctgttcccgtttt 240 ccataaaacc agatcttatc tccataaaca tcttttgtca tctgaacaaaatcaggccat 300 ttgcaaattg aatctataac ttctaaagga gcatctgaac gggaggccgaaaacgtatga 360 aaattaactg cccccaaacc aatatcactc atctccgcgt cttctag 40728 413 DNA Papilloma virus CDS (1)...(411) 28 att gag gac ggt gat atgatc gat att ggg ttt ggc aat ata aat aac 48 Ile Glu Asp Gly Asp Met IleAsp Ile Gly Phe Gly Asn Ile Asn Asn 1 5 10 15 aag aca tta tca gta aacaaa tca gat gtt agt tta gat tta gta aat 96 Lys Thr Leu Ser Val Asn LysSer Asp Val Ser Leu Asp Leu Val Asn 20 25 30 gaa ata gct aaa tat cca gatttt tta aca atg gct aat gat gtg tat 144 Glu Ile Ala Lys Tyr Pro Asp PheLeu Thr Met Ala Asn Asp Val Tyr 35 40 45 ggc gat tct tgc ttt ttt ttt gccagg aga gaa caa tgt tat gct aga 192 Gly Asp Ser Cys Phe Phe Phe Ala ArgArg Glu Gln Cys Tyr Ala Arg 50 55 60 cat tat ttt act aga gga gga gct gtgggt gat gct ata cct gat aca 240 His Tyr Phe Thr Arg Gly Gly Ala Val GlyAsp Ala Ile Pro Asp Thr 65 70 75 80 aca act aat caa gat cac aaa tac tatcta gca cct aag agt gga caa 288 Thr Thr Asn Gln Asp His Lys Tyr Tyr LeuAla Pro Lys Ser Gly Gln 85 90 95 tcc caa agt cct ttg ggt aat tct att tactat ccc acc gtt agt ggc 336 Ser Gln Ser Pro Leu Gly Asn Ser Ile Tyr TyrPro Thr Val Ser Gly 100 105 110 tcc tta gtt tct tct gat gca cag ctc tttaac aga ccc ttc tgg ttg 384 Ser Leu Val Ser Ser Asp Ala Gln Leu Phe AsnArg Pro Phe Trp Leu 115 120 125 aaa cgt gca cag ggg cac aat aac ggc at413 Lys Arg Ala Gln Gly His Asn Asn Gly 130 135 29 137 PRT Papillomavirus 29 Ile Glu Asp Gly Asp Met Ile Asp Ile Gly Phe Gly Asn Ile Asn Asn1 5 10 15 Lys Thr Leu Ser Val Asn Lys Ser Asp Val Ser Leu Asp Leu ValAsn 20 25 30 Glu Ile Ala Lys Tyr Pro Asp Phe Leu Thr Met Ala Asn Asp ValTyr 35 40 45 Gly Asp Ser Cys Phe Phe Phe Ala Arg Arg Glu Gln Cys Tyr AlaArg 50 55 60 His Tyr Phe Thr Arg Gly Gly Ala Val Gly Asp Ala Ile Pro AspThr 65 70 75 80 Thr Thr Asn Gln Asp His Lys Tyr Tyr Leu Ala Pro Lys SerGly Gln 85 90 95 Ser Gln Ser Pro Leu Gly Asn Ser Ile Tyr Tyr Pro Thr ValSer Gly 100 105 110 Ser Leu Val Ser Ser Asp Ala Gln Leu Phe Asn Arg ProPhe Trp Leu 115 120 125 Lys Arg Ala Gln Gly His Asn Asn Gly 130 135 30413 DNA Papilloma virus 30 atgccgttat tgtgcccctg tgcacgtttc aaccagaagggtctgttaaa gagctgtgca 60 tcagaagaaa ctaaggagcc actaacggtg ggatagtaaatagaattacc caaaggactt 120 tgggattgtc cactcttagg tgctagatag tatttgtgatcttgattagt tgttgtatca 180 ggtatagcat cacccacagc tcctcctcta gtaaaataatgtctagcata acattgttct 240 ctcctggcaa aaaaaaagca agaatcgcca tacacatcattagccattgt taaaaaatct 300 ggatatttag ctatttcatt tactaaatct aaactaacatctgatttgtt tactgataat 360 gtcttgttat ttatattgcc aaacccaata tcgatcatatcaccgtcctc aat 413 31 404 DNA Papilloma virus CDS (1)...(402) 31 ata caggat ggg gac atg ttt gat ata ggt ttt ggt aat att aac aat 48 Ile Gln AspGly Asp Met Phe Asp Ile Gly Phe Gly Asn Ile Asn Asn 1 5 10 15 aaa actcta tct tat aat aag tct gat gta agt tta gac att gtt aat 96 Lys Thr LeuSer Tyr Asn Lys Ser Asp Val Ser Leu Asp Ile Val Asn 20 25 30 gaa gta tgcaaa tat cca gat ttt ttg aca atg tct aat gat gtg tat 144 Glu Val Cys LysTyr Pro Asp Phe Leu Thr Met Ser Asn Asp Val Tyr 35 40 45 gga gac gca tgcttt tac tgt gcc cga aga gag caa tgt tat gct aga 192 Gly Asp Ala Cys PheTyr Cys Ala Arg Arg Glu Gln Cys Tyr Ala Arg 50 55 60 cat tat ttt gtt cgggga ggt cag gtt gga gat gca ata cct gac gag 240 His Tyr Phe Val Arg GlyGly Gln Val Gly Asp Ala Ile Pro Asp Glu 65 70 75 80 gca gtc caa caa gatcac aaa tat tat tta cct tct gat aca cgg cgc 288 Ala Val Gln Gln Asp HisLys Tyr Tyr Leu Pro Ser Asp Thr Arg Arg 85 90 95 act tta gaa aac tcc acctat ttt ccc acc gta agc ggg tcg ttg gtg 336 Thr Leu Glu Asn Ser Thr TyrPhe Pro Thr Val Ser Gly Ser Leu Val 100 105 110 acc tct gat gcc caa ctattt aat agg ccc ttt tgg tta aaa cgt gca 384 Thr Ser Asp Ala Gln Leu PheAsn Arg Pro Phe Trp Leu Lys Arg Ala 115 120 125 caa ggc cac aat aac ggaat 404 Gln Gly His Asn Asn Gly 130 32 134 PRT Papilloma virus 32 Ile GlnAsp Gly Asp Met Phe Asp Ile Gly Phe Gly Asn Ile Asn Asn 1 5 10 15 LysThr Leu Ser Tyr Asn Lys Ser Asp Val Ser Leu Asp Ile Val Asn 20 25 30 GluVal Cys Lys Tyr Pro Asp Phe Leu Thr Met Ser Asn Asp Val Tyr 35 40 45 GlyAsp Ala Cys Phe Tyr Cys Ala Arg Arg Glu Gln Cys Tyr Ala Arg 50 55 60 HisTyr Phe Val Arg Gly Gly Gln Val Gly Asp Ala Ile Pro Asp Glu 65 70 75 80Ala Val Gln Gln Asp His Lys Tyr Tyr Leu Pro Ser Asp Thr Arg Arg 85 90 95Thr Leu Glu Asn Ser Thr Tyr Phe Pro Thr Val Ser Gly Ser Leu Val 100 105110 Thr Ser Asp Ala Gln Leu Phe Asn Arg Pro Phe Trp Leu Lys Arg Ala 115120 125 Gln Gly His Asn Asn Gly 130 33 404 DNA Papilloma virus 33attccgttat tgtggccttg tgcacgtttt aaccaaaagg gcctattaaa tagttgggca 60tcagaggtca ccaacgaccc gcttacggtg ggaaaatagg tggagttttc taaagtgcgc 120cgtgtatcag aaggtaaata atatttgtga tcttgttgga ctgcctcgtc aggtattgca 180tctccaacct gacctccccg aacaaaataa tgtctagcat aacattgctc tcttcgggca 240cagtaaaagc atgcgtctcc atacacatca ttagacattg tcaaaaaatc tggatatttg 300catacttcat taacaatgtc taaacttaca tcagacttat tataagatag agttttattg 360ttaatattac caaaacctat atcaaacatg tccccatcct gtat 404

What is claimed is:
 1. An isolated polynucleotide consisting essentiallyof: (a) a nucleotide sequence encoding the peptide of SEQ ID NO:2, SEQID NO:5, or SEQ ID NO:8; (b) a nucleotide sequence hybridizing to thenucleotide sequence of SEQ ID NO:1, SEQ ID NO:4, or SEQ ID NO:7; or (c)the complement of (a) or (b); wherein the polynucleotide has a homologyof at least 90% to the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:4,or SEQ ID NO:7, or the complement thereof.
 2. An isolated polynucleotideencoding a peptide of a papilloma virus major capsid protein, whereinthe said polynucleotide has been obtained using the following steps: (a)incubating total DNA isolated from a biopsy of epithelial neoplasm witha nucleic acid having at least a portion of the nucleotide sequence ofSEQ ID NO:1, SEQ ID NO:4, or SEQ ID NO:7, under a condition that allowshybridization of a polynucleotide derived from a papilloma virus genomeincluded in the total DNA to said nucleotide sequence of the complementof SEQ ID NO:1, SEQ ID NO:4, or SEQ ID NO:7; and (b) identifying andisolating a polynucleotide that hybridizes to the complement of thenucleotide sequence of SEQ ID NO:1, SEQ ID NO:4, or SEQ ID NO:7 in step(a); wherein the polynucleotide has a homology of at least 90% to thenucleotide sequence of SEQ ID NO:1, SEQ ID NO:4, or SEQ ID NO:7.
 3. Anisolated polynucleotide, consisting essentially of (a) a nucleic acidencoding a peptide comprising the amino acid sequence of SEQ ID NO:2,SEQ ID NO:5, or SEQ ID NO:8, or (b) the complement of (a).
 4. Anisolated polynucleotide, wherein the polynucleotide consists essentiallyof the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:4, or SEQ ID NO:7,or the complement thereof.
 5. A plasmid comprising the polynucleotide ofclaim 1 or
 2. 6. A plasmid comprising the polynucleotide of claim 3 or4.
 7. An expression vector comprising the polynucleotide of claim 1 or2.
 8. An expression vector comprising the polynucleotide of claim 3 or4.
 9. A host cell comprising the plasmid of claim
 5. 10. A host cellcomprising the plasmid of claim
 6. 11. A host cell comprising theexpression vector of claim
 9. 12. A host cell comprising the expressionvector of claim
 8. 13. A method of producing a peptide of a papillomavirus major capsid protein, comprising cultivating the host cell ofclaim 11 under suitable conditions.
 14. A method of producing a peptideof a papilloma virus major capsid protein, comprising cultivating thehost cell of claim 12 under suitable conditions.
 15. A method ofdetecting a papilloma virus DNA, comprising: (a) hybridizing understringent conditions at least a portion of the polynucleotide of claim1, 2, 3, or 4 to a DNA sample; and (b) identifying papilloma virus insaid DNA sample by detecting a hybridization signal.
 16. A compositioncomprising the polynucleotide of claim 1, 2, 3, or 4 as reagent fordiagnosis and a diagnostically acceptable carrier.
 17. A method ofproducing a papilloma virus genome, comprising: (a) incubating total DNAisolated form a biopsy of epithlial neoplasm with a nucleic acid havingat least a portion of the nucleotide sequence of SEQ ID NO:1, SEQ IDNO:4, or SEQ ID NO:7, under a condition that allows hybridization of apolynucleotide derived from a papilloma virus genome included in thetotal DNA to said nucleotide sequence of the complement of SEQ ID NO:1,SEQ ID NO:4, or SEQ ID NO:7; and (b) identifying and isolating apolypnucleotide that hybridizes to the nucleotide sequence of thecomplement of SEQ ID NO:1, SEQ ID NO:4, or SEQ ID NO:7 in step (a). 18.The method of claim 17, wherein the polypnucleotide has a homology of atleast 90% to the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:4, or SEQID NO:7.
 19. A composition comprising the polynucleotide of claim 1, 2,3, or 4 as reagent for vaccination and a pharmaceutically acceptablecarrier.
 20. A method of vaccinating a subject in need against papillomavirus, comprising administering to said subject the composition of claim19.
 21. A method of diagnosing a condition caused by papilloma virus ina subject in need, comprising exposing said subject the composition ofclaim
 16. 22. A method of using the polynucleotide of claim 1, 2, 3, or4 as reagent for diagnosis.
 23. The method according to claim 22,wherein the diagnosis concerns papilloma virus infections or diseases.